圆斑星鲽雌性特异DNA分子标记开发及遗传性别鉴定方法的建立  

Identification of a female-specific DNA marker and development of genetic sex identification method in Verasper variegatus

作　　者：郑卫卫 徐文腾 刘洋[1] 陈亚东[1] 杨涛[1] 席晓晴 刘志鸿[1] 徐东[1] 秦搏[4] 陈松林[1,2] ZHENG Weiwei;XU Wenteng;LIU Yang;CHEN Yadong;YANG Tao;XI Xiaoqing;LIU Zhihong;XU Dong;QIN Bo;CHEN Songlin(State Key Laboratory of Mariculture Biobreeding and Sustainable Goods,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao Marine Science and Technology Center,Qingdao 266237,China;Rongcheng Marine Economic Development Center,Weihai 264300,China;East China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Shanghai 200090,China)

机构地区：[1]中国水产科学研究院黄海水产研究所,海水养殖生物育种与可持续产出全国重点实验室,山东青岛266071 [2]青岛海洋科技中心海洋渔业科学与食物产出过程功能实验室,山东青岛266237 [3]荣成市海洋经济发展中心,山东威海264300 [4]中国水产科学研究院东海水产研究所,上海200090

出　　处：《水产学报》2025年第5期20-28,共9页Journal of Fisheries of China

基　　金：科技创新2030—重大项目(2023ZD0405505);山东省重点研发计划项目(创新平台)(2022CXPT002);中央级公益性科研院所基本科研业务费专项(2023TD19);山东省泰山学者攀登计划;中国水产科学研究院东海水产研究所基本科研业务费专项(2016M08)。

摘　　要：【目的】开发能准确鉴定圆斑星鲽遗传性别的DNA分子标记。【方法】本研究首先通过全基因组重测序对圆斑星鲽雌性特异DNA序列进行了筛选,然后利用PCR扩增的方法对圆斑星鲽雌性特异DNA分子标记进行鉴定,并建立了一种圆斑星鲽遗传性别快速鉴定的方法。【结果】通过对17尾雄性和17尾雌性圆斑星鲽进行全基因组重测序,筛选出359条W染色体和Z染色体同源且W染色体特异性插入的差异DNA片段。根据差异DNA片段两翼保守序列设计雌雄共享引物对,采用PCR扩增和琼脂糖凝胶电泳方法鉴定出一个插入片段为69 bp的雌性特异DNA分子标记,使用该标记可在雌鱼中扩增出2条DNA条带,在雄鱼中扩增出1条DNA条带。进一步利用该DNA分子标记和PCR扩增方法对另一个养殖群体的117尾圆斑星鲽进行了遗传性别鉴定,鉴定结果与性腺组织切片结果一致。【结论】圆斑星鲽雌性特异DNA分子标记的开发及遗传性别鉴定方法的建立,不仅为圆斑星鲽性别决定机制研究提供了重要的遗传学证据,而且有助于加快圆斑星鲽性别控制育种和全雌苗种培育技术的发展和建立。Verasper variegatus,a marine flatfish,holds significant nutritional and economic value and exhibits sexual dimorphism in growth,with females growing faster than the males.However,to date,no rapid,effective,and universal sex-specific DNA marker has been available for genetic sex identification and sex-controlled breeding in V.variegatus.To address this,we performed whole genome resequencing on 17 male and 17 female individuals to identify female-specific DNA sequences.Through PCR amplification,we identified a female-specific DNA marker and established a genetic sex identification method.Our results revealed 359 potential female-specific DNA sequences following reads mapping,sequencing depth,and coverage analysis.Primer pairs targeting approximately 150 bp DNA sequences flanking these regions were designed.After two rounds of PCR verification,a 69 bp insertion was identified as a female-specific marker,accurately distinguishing genetic sex via PCR amplification and 2%agarose gel electrophoresis.Further validated using 117 individuals from another breeding population confirmed the accuracy of this marker and method,with genetic sex results aligning with phenotypic determined by gonad histology.The development of this female-specific DNA marker and the genetic sex identification method provides crucial genetic insights into the sex determination mechanism of V.variegatus and accelerates the implementation of sex-controlled breeding and all-female breeding techniques.

关 键 词：圆斑星鲽 雌性特异DNA分子标记 遗传性别鉴定 基因组重测序 

分 类 号：S917.4[农业科学—水产科学]